Hello.
I'm making non-denaturing PAGE of DNA fragments for analysis of polymorphisms, but I'm getting wavy bands. If somebody can help me, I will be very greatful.
First of all, here's my recipe for a total volume of 10 mL for a 12% polyacrylamide gel.
H2O 4.95 mL
30% acrylamide mix 4.00 mL
TBE 5X 1.00 mL
APS 50 uL
TEMED 10 uL
After casting the gel, I let it polymerize for 90 minutes.
Then, before charging, I make sure to rinse all the wells.
After that, I charge 3uL of my PCR products and 3.5uL of my DNA ladder.
For the electrophoresis buffer I use 0.5X TBE.
This gel in the image was run at 85V. I also made another run at 40V, but I had the same result of wavy bands.
I hope somebody could help me if I'm making mistakes or if I can do something better. Thank you so much!
2 things are worth trying. Wash the comb thoroughly and clean it well. Wash out the loading wells with a pipette or syringe immediately before loading the samples
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